In this renewal, the RESBIO Biomedical Technology Resource Center will build on its pioneering achievements in the development and application of advanced methods for discovery and optimization of biomaterials. The goals of the third funding period are a) expanding understanding of basic signaling mechanisms involved in cell-materials interactions, b) advancing new methods and technologies for creation of tissue scaffolds, and c) testing these advances in applications for stem cell research, tissue engineering and regenerative medicine. Six highly coordinated Technology Research and Development Projects will be conducted by a team spanning five universities and one national laboratory. In Project 1, chemists and material scientists will prepare hydrogels with ligands expressed throughout the volume and nanofiber mats with surface-tethered ligands to facilitate the study of specific biological interactions. They will also fabricate defined test beds that can be distributed among other Core projects, collaborators, and the biomedical community. In Project 2, biologists will develop composite biologic-synthetic materials that support stem cell self-renewal. Project 3 will develop approaches to enable the study of how mechanical interactions between cells and biomaterials regulate cell function and thus inform the design and fabrication of bioactive scaffolds. The Project 4 team will advance imaging-based methods to study biomaterial-driven organization of nuclear and cytoskeletal probes used to characterize stem cell intracellular signaling and organizational behaviors in 3D environments. Project 5's goal is to develop a noninvasive functional Raman imaging tool to produce 3D, chemically resolved images of live cells and tissues at microscale resolution. Project 6 will extend its multiscale computational molecular modeling approaches to guide optimization of the bioactive polymer design in Projects 1 and 2. Collaborations will challenge RESBIO's emerging technologies with projects on stem cell characterization; bone, soft tissue, and nerve regeneration; discovery of angiogenic biomaterials; and protein-polymer interactions. Well-established programs of Service, Training and Dissemination will be expanded with new content from the Core Projects, and delivered to a wider scientific community.